Gibson D. (2009) Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides. Nucleic Acids Res, 37:6984–6990.
When producing large, synthetic genes it is frequently necessary to join many chemically synthesized DNA elements, which is often impossible with traditional blunt- and cohesive-end cloning methods. Dr Dan Gibson, at Synthetic Genomics Inc., is a pioneer in developing methods for assembling genes from pools of synthetic, single- and double-stranded DNA. In this article, Dr Gibson describes an interesting protocol for assembling high-fidelity Ultramer™ oligonucleotides
with overlapping sequences into large constructs using Saccharomyces cerevisiae
. The method can be used for both single- and double-stranded DNA, including IDT gBlocks™ Gene Fragments
. In a recent article, Isothermal Assembly: Quick, Easy Gene Construction
, available at www.idtdna.com/DECODED, we discussed the more recently developed Gibson Assembly™ Method, which uses a unique enzyme mix to rapidly assemble large synthetic genes.